Atractylodin ameliorates obesity-associated hepatic steatosis by regulating the PLIN2-ATGL/CPT1A axis-mediated lipid droplet-mitochondria interactions

Background Obesity prevalence is rising, increasing risks of ‌metabolic associated fatty liver disease, diabetes mellitus, and cardiovascular disease. Atractylodin (ATR), a bioactive compound from Atractylodes lancea , exhibits potential against metabolic disorders, but its mechanisms against obesity-driven hepatic steatosis remain unclear. Purpose This study aimed to investigate the therapeutic effects and underlying mechanisms of ATR in ameliorating obesity-associated hepatic steatosis via lipid droplet (LD)-mitochondria interactions. Methods Mice were fed a high-fat diet (HFD) for 12 weeks, followed by administration of ATR or Orlistat for 6 weeks beginning at week 6. Hepatic steatosis and molecular targets were assessed via serum biochemistry, histology, fluorescent staining, and Western blot. AML-12 and HepG2 cells were induced with free fatty acids (FFA) and treated with ATR, Atglistatin (ATGLi), etomoxir (ETO), subjected to CPT1A knockdown, or subjected to PLIN2 overexpression/knockdown. In addition to analyzing key proteins and lipid content by molecular and biochemical methods, LD-mitochondria contacts were visualized by transmission electron microscope and laser confocal fluorescence microscopy. Results ATR significantly ameliorated obesity-associated hepatic steatosis in mice and reduced lipid accumulation in AML-12 and HepG2 cells. Visual analysis confirmed that ATR promoted LD-mitochondria contacts. Molecular analysis showed that ATR regulated proteins for lipolysis and fatty acid oxidation (FAO) in liver. Genetic approaches validated that ATR downregulated PLIN2. Mechanistically, ATR promoted LD-mitochondria interactions, lipolysis, and FAO. The effects may be mediated through the regulation of the PLIN2-ATGL/CPT1A axis. Conclusions ATR ameliorates obesity-associated hepatic steatosis by enhancing lipolysis and FAO. Promoting LD-mitochondria interactions represents a promising therapeutic strategy, with ATR showing potential as a therapeutic agent for obesity.